U.S. patent application number 12/162326 was filed with the patent office on 2009-12-10 for therapeutic indications of colony stimulating factors.
This patent application is currently assigned to SYGNIS Bioscience GmbH & Co., KG. Invention is credited to Alfred Bach, Hans-Jurgen Quadbeck-Seeger.
Application Number | 20090305974 12/162326 |
Document ID | / |
Family ID | 38255129 |
Filed Date | 2009-12-10 |
United States Patent
Application |
20090305974 |
Kind Code |
A1 |
Bach; Alfred ; et
al. |
December 10, 2009 |
THERAPEUTIC INDICATIONS OF COLONY STIMULATING FACTORS
Abstract
The present invention relates to the use of at least one colony
stimulating factor (CSF) for the production of medicinal products
in the treatment or prophylaxis of coma or neurotoxicity.
Inventors: |
Bach; Alfred;
(Edingen-Neckarhausen, DE) ; Quadbeck-Seeger;
Hans-Jurgen; (Bad Durkheim, DE) |
Correspondence
Address: |
FOLEY AND LARDNER LLP;SUITE 500
3000 K STREET NW
WASHINGTON
DC
20007
US
|
Assignee: |
SYGNIS Bioscience GmbH & Co.,
KG
|
Family ID: |
38255129 |
Appl. No.: |
12/162326 |
Filed: |
January 24, 2007 |
PCT Filed: |
January 24, 2007 |
PCT NO: |
PCT/EP07/50685 |
371 Date: |
March 13, 2009 |
Current U.S.
Class: |
514/7.5 |
Current CPC
Class: |
A61P 25/02 20180101;
A61K 38/193 20130101; A61P 25/00 20180101 |
Class at
Publication: |
514/12 |
International
Class: |
A61K 38/18 20060101
A61K038/18; A61P 25/00 20060101 A61P025/00 |
Foreign Application Data
Date |
Code |
Application Number |
Jan 27, 2006 |
DE |
10 2006 004 142.9 |
Claims
1.-8. (canceled)
9. A method for the treatment or prophylaxis of neurotoxicity,
wherein a therapeutically effective dose of G-CSF, GM-CSF or of a
polypeptide with an identity of at least 90% to either G-CSF or
GM-CSF is administered to a patient in need thereof.
10. The method as claimed in claim 9, wherein the G-CSF is human
G-CSF.
11. The method as claimed in claim 9, wherein the GM-CSF is human
GM-CSF.
12. The method as claimed in claim 9, wherein the neurotoxicity is
a neurotransmission-associated neurotoxicity.
13. The method as claimed in claim 9, wherein the neurotoxicity is
a neurotoxicity induced by amphetamine or derivatives thereof.
14. The method as claimed in claim 13, wherein the amphetamine
derivative is a 3,4-methylenedioxymethylamphetamine.
Description
[0001] The colony stimulating factors (CSF) are a group of
regulatory proteins that are responsible for controlling the
proliferation and differentiation of hematopoietic cells such as
granulocytes, megakaryocytes and monocytes or macrophages. Without
appropriate CSFs, these hematopoietic cells cannot survive and/or
proliferate in culture. The CSFs belong to the cytokine group.
Together with erythropoietin (EPO) and some interleukins, they form
the group of hematopoietic growth factors.
[0002] The CSF group includes the factors M-CSF (macrophage-colony
stimulating factor; also CSF-1), GM-CSF
(macrophage/granulocyte-colony stimulating factor; also CSF-2),
G-CSF (granulocyte-colony stimulating factor; also CSF-3) and
multi-CSF (multifunctional colony stimulating factor; also IL3)
according to their specificity with respect to the various
hematopoietic cells. Purification and cloning of the individual
CSFs makes molecular characterization possible. The aforesaid four
CSFs are glycoproteins, but they do not display any homology at the
level of the primary structure (amino acid sequence).
[0003] M-CSF is produced by monocytes, granulocytes, endothelial
cells and fibroblasts. Furthermore, however, activated B- and
T-cells, as well as a number of tumor cell lines are able to
synthesize this factor. M-CSF is a homodimeric glycoprotein. The
sugar moiety is not necessary for biological activity. There are
several variants with different molecular weights, which result
from alternative splicing of the RNA.
[0004] M-CSF promotes the proliferation and differentiation of
hematopoietic stem cells to macrophages, but mainly the growth,
differentiation and functional activity of monocytes. Whereas human
M-CSF also exhibits activity in mouse and rat cells, the murine
factor is inactive in human cells.
[0005] G-CSF is secreted by activated monocytes, macrophages and
neutrophils, by stroma cells, fibroblasts and endothelial cells,
and by various tumor cell lines (e.g. human bladder cancer cell
line). Mature human G-CSF is a monomeric glycoprotein with 174
amino acids, where the sugar moiety is not necessary for biological
activity. Another variant with 177 amino acids, resulting from
alternative splicing of the RNA, displays greatly reduced
biological activity.
[0006] G-CSF promotes the proliferation and differentiation of
hematopoietic precursor cells to neutrophilic granulocytes and also
activates these. Furthermore, G-CSF also acts as a mitogenic
agent.
[0007] The most important clinical application of G-CSF is the
treatment of leukopenia, e.g. following chemotherapy and/or
radiotherapy.
[0008] GM-CSF is a monomeric glycoprotein of 127 amino acids, where
the sugar moiety is not necessary for biological activity. The
GM-CSF receptor occurs not only on hematopoietic cells, but also
e.g. on endothelial cells.
[0009] The specificity of GM-CSF is generally less pronounced than,
for example, that of G-CSF. Thus, GM-CSF stimulates the
proliferation and differentiation of neutrophil, eosinophil and
monocyte lines and activates their mature form. At low
concentrations the factor exerts a chemotactic action on
eosinophils and neutrophils. As GM-CSF is produced by the cells
(T-lymphocytes, macrophages, endothelial cells and mast cells) that
are involved in an inflammatory response, it can be assumed that
this factor plays an important role as mediator in
inflammation.
[0010] In synergy with EPO, GM-CSF also promotes the proliferation
of erythroid and megakaryocyte precursor cells.
[0011] GM-CSF finds clinical application for reconstitution of
hematopoiesis. Its most important use is for the treatment of
neutropenia, e.g. in connection with chemotherapy or
radiotherapy.
[0012] Multi-CSF is mainly produced by activated T-cells, but also
by keratinocytes, NK cells, mast cells, endothelial cells and
monocytes. Mature human multi-CSF is a glycoprotein of 133 amino
acids, where the sugar moiety is not necessary for biological
activity.
[0013] Multi-CSF has a very broad spectrum of biological
activities. Thus, multi-CSF supports the proliferation and
differentiation of nearly all types of hematopoietic precursor
cells. As initial factor, it makes the hematopoietic stem cells
responsive to later-acting factors such as EPO and GM-CSF. The
biological activities of multi-CSF are species-specific.
[0014] In view of this promotion of proliferation, differentiation
and activation of cells of the hematopoietic system, CSFs are used
therapeutically for reconstitution of hematopoiesis. Accordingly,
mainly recombinant G-CSF (e.g. filgrastim) is used therapeutically
for the treatment of neutropenia following chemotherapy or
radiotherapy.
[0015] Furthermore, yet more therapeutic applications are described
for CSFs. In this connection we may mention e.g. the use of CSF for
the treatment of infections (WO 88/00832) and for promoting wound
healing (WO 92/14480). The observation that CSFs may also play an
important role in angiogenesis (WO 97/14307) and particularly in
arteriogenesis (WO 99/17798) offers the possibility of using these
factors for the treatment of ischemias such as myocardial
infarction and stroke, by restoring and/or improving the blood flow
in the affected tissue.
[0016] It has also been observed that certain CSF receptors are
also present on neurons (DE-A 100 33 219). Accordingly, a
neuroprotective and neuroregenerative action was recently
demonstrated for G-CSF for the treatment of focal cerebral ischemia
in an animal model [Schabitz et al. Stroke. 34:745 (2003);
Schneider et al. J Clin Invest. 115:2083 (2005)].
[0017] A condition for which there continues to be a large demand
for suitable medicinal products is coma. Coma is a severe degree of
disturbance of consciousness, in which the patient can no longer be
woken by external stimuli (Pschyrembel, 259th Edition, 2002; pp.
603-604, pp. 882-883, p. 978, p. 1110, p. 1620). Depending on the
symptoms and the causes, the following coma states are
distinguished, among others:
[0018] Apallic syndrome (persistent vegetative state): a clinical
picture that is included among the decerebration syndromes, with
functional loss of the cerebral cortex, generally as a result of
anoxia of the brain (e.g. after head injury, intoxication, shock or
resuscitation) and disturbance of the ascending reticular
activating system with retention of brainstem function. The patient
is awake and his eyes are open, but he does not show any
spontaneous and reactive movements and also no eye fixation. Also
there are no spontaneous utterances. Spontaneous breathing and
circulatory regulation are, however, intact. If caused by injury or
infection, functional recovery is still possible after some months,
though it is unlikely after more than three months. In the absence
of remission, death occurs after two to five years (e.g. as a
result of complications such as pneumonia, urinary tract infection
or decubitus).
[0019] "Locked-in" syndrome: inability, although remaining
conscious, to communicate spontaneously by speech or through
movements. Communication through eye movements is possible. The
cause is a bilateral transverse lesion of the tractus
corticobulbaris and tractus corticospinalis in the region of the
pons, e.g. in arteria basilaris thrombosis. The prognosis is
unfavorable.
[0020] Akinetic mutism: mutism (dumbness) as a result of general
inhibition of motor capabilities including facial expression,
gestures and speech. Speech and movement do not take place
spontaneously, and only slowly and with a delay after being
requested. There is also disturbance of the sleeping-waking cycle.
Pain stimuli increase vigilance and make limited contact possible.
Consciousness is fully retained, and there may be amnesia. Akinetic
mutism occurs e.g. after frontal lobe lesions, in psychoses, tumor
or hemangioma near the 3rd ventricle of the brain or in the
mesencephalon in arteria basilaris thrombosis or in
encephalitis.
[0021] The Glasgow Coma Scale (GCS) permits quantitative
classification according to the severity of the disturbance of
consciousness ("mild disturbance of consciousness" (14-15 points);
"moderate disturbance of consciousness" (13-9) points; "severe
disturbance of consciousness" (3-8 points)). The patient's reaction
is assessed in three areas (eye opening, motor behavior and speech)
and the corresponding points achieved are added together.
[0022] Treatment of coma patients should mainly be directed towards
increasing the probability of awakening and reducing or reversing
the neurological damage caused by being in the comatose state.
Initial studies show such a therapeutic effect e.g. for treatment
with acetyl-L-carnitine (EP-A 0 498 144). At present, however, no
commercial preparation that fulfils this therapeutic requirement is
in clinical use.
[0023] Owing to their particular properties, neurons have increased
sensitivity to the action of toxic substances (Anthony, Montine,
Valentine & Graham; Toxicology; Ed. Casarett & Doull; 6th
edition, 2001; pp. 535-563).
[0024] Thus, because of their high energy requirements, neurons are
particularly dependent on aerobic metabolism. Even short
interruptions in the supply of oxygen or glucose can damage the
neurons. An example of this is hypoxia as a result of carbon
monoxide poisoning, in which there is mainly damage to the neurons
that are especially sensitive to this, such as those in certain
regions of the cerebral cortex.
[0025] Another reason for the particular sensitivity of the neurons
to toxic substances is the typical structure of the neurons with
their long processes, the axons. These axons, the length of which
can reach 200 000 times the diameter of the cell body, must be
supplied from the cell body. The provision of protein synthesis
machinery for such a large volume of cytoplasm and the axonal
transport of the synthesis products make heavy demands on the
neurons.
[0026] The special sensitivity of the cells of the nervous system
is countered by a special protection, the blood-brain barrier. In
contrast to the blood vessels of other organs, the cerebral
capillary vessels are enveloped by their epithelial cells without
any gaps ("tight junctions"). In addition, the surrounding glial
cells form a further barrier to the passive transport of many
substances as well as toxins. The substances that the nerve cells
require from the blood are brought in through the barrier by active
transport. Only lipid-soluble substances, and hence also toxins of
that type, can penetrate the blood-brain barrier passively. The
protection afforded by the blood-brain barrier has some other
important limitations. Thus, certain regions of the nervous system,
e.g. the circumventricular organ, are not protected by a
blood-brain barrier. Moreover, the blood-brain barrier of neonates
is not yet fully manifested.
[0027] Depending on the mechanism of neurotoxicity, a distinction
is made between four different subgroups: neuronopathy, axonopathy,
myelinopathy and transmission-associated toxicity.
[0028] A neuronopathy means there is primarily a lesion of the
neuron cell body. The loss of a neuron is irreversible and includes
the degradation of all cytoplasmic processes, such as dendrites and
axons, and of the associated myelin. Various neurotoxins are
specific for certain neuronal subpopulations and so can lead to
characteristic losses of function. Doxorubicin for example, a
cytostatic, which is deposited in the DNA double strand, mainly
damages the neurons of the peripheral nervous system (PNS) and
other nerve cells not protected by the blood-brain barrier. The
neurotoxin MPTP (1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine) can,
in contrast, owing to its uncharged state, penetrate the
blood-brain barrier. Then it is oxidized enzymatically to the
corresponding pyridinium ion (MPP) and taken up via the
dopamine-transport system mainly by the dopaminergic neurons of the
substantia nigra. There, the MPP blocks the mitochondrial cellular
respiration and so leads to the death of these neurons. The
symptoms of this intoxication correspond to those of irreversible
Parkinson's disease. Low doses of MPTP, which do not cause any
acute symptoms, can increase the predisposition for Parkinson's
disease. Other neurotoxins acting by the mechanism of neuronopathy
include some heavy metals (e.g. lead, bismuth, mercury and
manganese), some antibiotics (e.g. chloramphenicol and
streptomycin) and alcohols (e.g. methanol and ethanol).
[0029] In the case of the neurotoxic diseases that come under the
heading axonopathy, primarily the axon is damaged. Following the
primary damage, generally there is degradation of the distal end of
the axon in a multistage process, whereas the cell body of the
neuron survives. Whereas this degradation in the central nervous
system is irreversible, the axons of the peripheral nervous system
can regenerate. n-Hexane and carbon disulfide are examples of
neurotoxins that act in this way. They lead to crosslinking of the
axonal neurofilaments, followed by swelling of the axons and
impairment of neurofilament transport. Generally axonopathy leads
to peripheral neuropathy. The sensory and motor impulse conduction
is increasingly impaired. In contrast, acrylamide-induced
axonopathy begins with degeneration of the distal end of the axon
in conjunction with damage to retrograde transport. The primary
point of action of some other neurotoxins such as colchicines and
paclitaxel (Taxol) is microtubule-based transport along the axons.
Taxol binds to the tubuli and colchicine binds to monomeric
tubulin. In this way they disturb the dynamic equilibrium of
formation and degradation of the microtubules.
[0030] Myelinopathies are neurotoxic diseases based on damage to
myelin. Myelin, which is formed by the oligodendrocytes in the
central nervous system (CNS) and by the Schwann cells in the
peripheral nervous system (PNS), is necessary for efficient impulse
conduction along the axons. Whereas the Schwann cells of the PNS
make it possible for myelin to be regenerated after neurotoxic
damage, remyelination is only possible to a limited extent in the
CNS. Hexachlorophene, for example, binds firmly to cell membranes
and leads to a loss of the ion gradient and finally to edema
between the myelin layers. The symptoms of acute hexachlorophene
intoxication start with general weakness, irritations and spasms,
ultimately leading to coma and death.
[0031] In neurotransmission-associated neurotoxicity, primarily the
process of neurotransmission is impaired. The neurotoxins of this
subclass interrupt impulse conduction, block or intensify
trans-synaptic communication, block reuptake of the
neurotransmitter or interfere with the "second messenger" system.
In most cases these neurotoxins display short-term, reversible
interactions, which subside after acute exposure, or can be
counteracted with suitable antagonists. In chronic exposure,
however, there may also be irreversible long-term consequences.
Examples of some neurotoxins of this subgroup are given below.
[0032] Nicotine, for example, binds agonistically to certain
cholinergic receptors. Small doses of nicotine lead to accelerated
heartbeat, raised blood pressure and narrowing of peripheral blood
vessels. Acute poisoning with nicotine results in sudden
overstimulation of the nicotinergic receptors followed by paralysis
of the ganglia, which can lead to respiratory arrest.
[0033] The euphoriant and habit-forming property of cocaine can be
attributed to changes in catecholaminergic neurotransmission. It is
mainly dopaminergic neurotransmission that is intensified by
blocking of the "dopamine reuptake transporter". Cocaine abuse is
associated with increased risk of cerebrovascular diseases,
cerebral perfusion defects and cerebral atrophy. Chronic cocaine
consumption is apparently associated with neurodegenerative changes
in the striatum, which is probably the cause of the neurological
and psychiatric symptoms.
[0034] Like cocaine, the amphetamines also affect catecholaminergic
neurotransmission. The neurotoxicity of the amphetamine derivative
3,4-methylenedioxymethylamphetamine has been much debated recently
(MDMA, "Ecstasy") (R. Mathias, NIDA Notes Vol. 14, No. 4 "Ecstasy
damages the brain and impairs memory in humans" (1999)). This drug
stimulates the release of serotonin and leads to a psychedelic
state. In addition, the need for food, drink and sleep is
suppressed. The acute toxic effects of MDMA include nausea, chills,
hallucinations, raised body temperature, trembling, muscle cramps
and blurred vision. An overdose leads to high blood pressure,
tiredness and panic attacks, and in more severe cases to
unconsciousness, convulsions and dramatically raised body
temperature. Cardiac failure and heat stroke may develop as a
result of an overdose. As well as this acute neurotoxicity,
however, it can be assumed there will also be chronic neurotoxic
sequelae. There are indications that regular use of MDMA leads to
damage of the serotonin-releasing neurons. This is accompanied by
significant impairment of memory. There is probably also
disturbance of other serotonin-dependent brain functions, such as
mood and the sleep cycle. Animal experiments suggest that the
damage to the neurons lasts many years, and may even be
permanent.
[0035] The aim of the present invention is to provide novel,
effective medicinal products for the treatment of coma and/or
neurotoxicity.
[0036] This aim is achieved through the use of at least one colony
stimulating factor for the production of a medicinal product for
the treatment or prophylaxis of coma and/or neurotoxicity.
[0037] Preferably this CSF is a colony stimulating factor from the
group G-CSF, M-CSF and GM-CSF. Especially preferably, G-CSF and/or
GM-CSF are used as the therapeutic active substance. In particular,
human polypeptides, i.e. human G-CSF and/or GM-CSF in the form of
recombinant proteins, are used as the therapeutic active
substances.
[0038] The term "colony stimulating factor" also includes variants
of the aforementioned specific factors. These can be homologous,
orthologous or paralogous sequences. Said variants include
sequences that have at least one base substitution, a base addition
or a base deletion, and the variants should always be a polypeptide
with the aforementioned biological activity of the respective
starting sequence. Functionally homologous variants and derivatives
are also included, e.g. PEGylated polypeptides or polypeptides for
which the activity of the colony stimulating factors has been
improved or prolonged in some other manner.
[0039] Polypeptides that are at least 70%, at least 75%, at least
80%, at least 81%, at least 82%, at least 83%, at least 84%, at
least 85%, at least 86%, at least 87%, at least 88%, at least 89%,
at least 90%, at least 91%, at least 92%, at least 93%, at least
94%, at least 95%, at least 96%, at least 97%, at least 98% or at
least 99%, or to some other percentage stated herein, identical to
an amino acid sequence of one of the aforementioned specific colony
stimulating factors and where the polypeptide has the respective
biological activity of the starting sequence, are also
included.
[0040] The percentage of identical amino acids preferably relates
to a sequence segment of at least 50% of the sequences to be
compared and especially preferably over the entire length of the
sequences to be compared. A large number of programs that implement
algorithms for said comparisons are described in the prior art and
are commercially available. Reference may be made in particular to
the algorithms of Needleman and Wunsch or Smith and Waterman, which
provide especially reliable results. These algorithms can
preferably be implemented by means of the following programs:
PileUp (J. Mol. Evolution., 25, 351-360, 1987, Higgins et al.,
CABIOS, 1989: 151-153), Gap and BestFit (Needleman and Wunsch (J.
Mol. Biol. 48; 443-453 (1970)) and Smith and Waterman (Adv. Appl.
Math. 2: 482-489 (1981))), as part of the GCG software [Genetics
Computer Group, 575 Science Drive, Madison, Wis., USA 53711
(1991)]. Especially preferably, the percentage (%) of sequence
identity is determined, within the scope of the present invention,
with the GAP program over the complete sequence with the following
established values: Gap Weight: 50, Length Weight 3, Average Match:
10.000 and Average Mismatch: 0.000.
[0041] A polypeptide that only comprises a fragment of the
aforementioned colony stimulating factors is also a polypeptide
according to the invention. The fragment should then encode a
polypeptide that has the biological activity of the starting
polypeptide. Said polypeptides comprise or therefore consist of
domains of the aforementioned specific polypeptides (starting
polypeptides), which impart the biological activity. A fragment in
the sense of the invention preferably comprises at least 20, at
least 30, at least 50, at least 80, at least 100 or at least 150
successive amino acids of an amino acid sequence of one of the
aforementioned colony stimulating factors.
[0042] The variants according to the invention preferably have at
least 10%, at least 20%, at least 30%, at least 40%, at least 50%,
at least 60%, at least 70%, at least 80% or at least 90% of the
respective biological activity of the starting polypeptide. The
variants according to the invention can, however, also have
improved activity relative to the starting polypeptide.
[0043] The invention relates moreover to the preferred use
according to the invention of colony stimulating factors for the
production of a medicinal product for the treatment of the coma
states apallic syndrome, "locked-in" syndrome and akinetic
mutism.
[0044] A further object of the invention comprises the use of
colony stimulating factors for the production of a medicinal
product for the treatment of neurotransmission-associated
neurotoxicity. In particular, said neurotoxicity can have been
induced by amphetamine or its derivatives (in particular
3,4-methylenedioxymethylamphetamine).
[0045] The CSFs can be administered in a variety of forms,
including, among others: solutions for infusion or injection,
suspensions, tablets, pills, powders, sprays or suppositories. The
preferred form depends on the method of administration and the
therapeutic application. The method of administration can be, among
others: oral, subcutaneous, pulmonary, intranasal, intramuscular,
rectal, intracerebral or intravenous administration. The preferred
method of administration is intravenous injection or infusion.
[0046] The therapeutically effective dose of the CSFs, which can be
administered either alone or as a combination of various CSFs,
should be chosen so that a neuroprotective effect is achieved.
Therefore the dose can in particular be in a range between 100 ng
and 10 mg/kg body weight. By taking into account factors such as
the patient's age, sex and severity of the neurological disturbance
and selection of the CSF or CSFs used can lead to individually
tailored doses. A further modification of the dose can follow from
the method of administration and the associated pharmacokinetics
and local availability. For example, the dose would be lower in the
case of direct intracerebral injection. In certain cases of
treatment of the neurological disorders described here, the use of
high doses of CSF (e.g. more than 1 mg/kg body weight) can be
especially useful.
[0047] The treatment is preferably started within the first week
after onset of the neurological disturbance, but later commencement
of treatment is also possible with these often chronic neurological
disorders. For treating said chronic forms of neurological
disorders, a regular, preferably daily, dose of CSF can be
administered. It can then preferably be administered in a
formulation that permits slow, continuous release of the active
substance ("slow-release formulation"). This slow, continuous
administration of the active substance can also be achieved e.g. by
infusion or using micrometering pumps.
[0048] The pharmacological preparations with one or more CSFs as
active substance can be prepared by the standard methods from the
prior art that are known by a person skilled in the art.
Pharmaceutically acceptable excipients can be added to the
preparation. The appropriate form of the pharmacological
preparation and the method of administration can be selected in
relation to the neurological disturbance to be treated, its
severity and other relevant circumstances. The pharmacological
preparation can be adapted for oral, parenteral or topical
administration. The CSFs used as the active substance can also be
used in the form of a pharmacologically acceptable salt, e.g. for
reasons of stability, solubility or better crystallizability.
* * * * *